Graziotto Romina, Del Prete Dorella, Rigotti Paolo, Anglani Franca, Baldan Nicola, Furian Lucrezia, Valente Marialuisa, Antonello Augusto, Marchini Francesco, D'Angelo Angela, Gambaro Giovanni
Clinic of Nephrology, Department of Medical and Surgical Science, University of Padova, Padova, Italy.
Transplantation. 2006 Apr 27;81(8):1125-32. doi: 10.1097/01.tp.0000208573.16839.67.
The Perforin-Granzyme B and Fas/Fas Ligand apoptotic mechanisms are involved in the development of acute renal rejection (AR). We describe our experience of analyzing the expression of cytotoxic T-lymphotoxins (CTL) in biopsies and peripheral blood leukocytes (PBL) for the diagnosis of AR.
We analyzed Perforin (P), Granzyme B (GB) and Fas Ligand (FL) expression in 68 renal biopsies and 64 PBL using comparative kinetic RT-PCR and, for GAPDH and FL, we also replicated with real-time RT-PCR. The levels of expression were measured in different groups, such as T0 (biopsies before reperfusion and PBL in recipient before the transplant [Tx]), Td (biopsies and PBL collected for clinical purposes) and Tp (biopsies and PBL two months after Tx).
A higher CTL expression was seen in non-rejecting (NR) biopsies in the first 2 months after Tx. P and FL were significantly more expressed during AR in all biopsies and in Td, while P remained upregulated in Tp. In PBL, there was no significant increase in CTL transcription during AR. A variable expression of CTL emerged in all T0 biopsies.
Two lytic pathways are activated in biopsies when AR occurs shortly after Tx, whereas the P/GB mechanism prevails if it occurs later on. Only P and FL in biopsies might be able to predict AR diagnosis, but with a considerable variability in each sample, possibly due to the small portion of tissue core, which may be inadequate for molecular diagnosis. CTL expression in PBL does not correlate with histological AR.
穿孔素 - 颗粒酶B和Fas/Fas配体凋亡机制参与急性肾移植排斥反应（AR）的发生发展。我们描述了分析活检组织和外周血白细胞（PBL）中细胞毒性T淋巴细胞毒素（CTL）表达以诊断AR的经验。
我们使用比较动力学逆转录聚合酶链反应（RT-PCR）分析了68例肾活检组织和64份PBL中的穿孔素（P）、颗粒酶B（GB）和Fas配体（FL）表达，对于甘油醛 - 3 - 磷酸脱氢酶（GAPDH）和FL，我们还采用实时RT-PCR进行了重复检测。在不同组中测量表达水平，如T0（再灌注前的活检组织以及移植前（Tx）受体的PBL）、Td（为临床目的采集的活检组织和PBL）和Tp（Tx后两个月的活检组织和PBL）。
Tx后前2个月，在无排斥反应（NR）的活检组织中观察到较高的CTL表达。在所有活检组织和Td组中，AR期间P和FL的表达明显增加，而P在Tp组中仍上调。在PBL中，AR期间CTL转录没有显著增加。在所有T0活检组织中出现了CTL的可变表达。
Tx后不久发生AR时，活检组织中有两条溶解途径被激活，而如果AR稍后发生，则P/GB机制占主导。活检组织中只有P和FL可能能够预测AR诊断，但每个样本存在相当大的变异性，可能是由于组织芯的部分较小，可能不足以进行分子诊断。PBL中的CTL表达与组织学AR不相关。
